Class III beta-tubulin isotype (betaIII-tubulin) is widely regarded as a neuronal marker in developmental neurobiology and stem cell research. To test the specificity of this protein, we determined its expression distribution primary cultures glial fibrillary acidic protein (GFAP)-expressing astrocytes isolated from cerebral hemispheres 2 human fetuses at 18 to 20 weeks gestation. Cells were maintained monolayer for 1 21 days without differentiation induction. By immunofluorescence...

Mast cell activation mediated by the high affinity receptor for IgE (FcεRI) is a key event in allergic response and inflammation. Other receptors on mast cells, as c-Kit stem factor G protein-coupled (GPCRs) synergistically enhance FcεRI-mediated release of inflammatory mediators. Activation various signaling pathways cells results changes morphology, adhesion to substrate, exocytosis, migration. Reorganization cytoskeleton pivotal all these processes. Cytoskeletal proteins also play an...

ABSTRACT To investigate the effects of heat stress on plant cytoskeleton, structure microtubule arrays in N. tabacum suspension cells incubated at 38 or 42°C was analysed. Whilst incubation 42 °C resulted disruption majority cellular microtubules after 30 min, exposed to all were preserved even 12 h incubation, although their organization altered. The most susceptible mitotic spindle and phragmoplast. Several abnormalities observed: (i) splitting into several parts; (ii) elongation spindles;...

Abstract Glial fibrillary acidic protein, GFAP, is a major intermediate filament protein of glial cells and cytoskeletal structure in astrocytes. The entorhinal cortex has key role memory function one the first brain areas to reveal hallmark structures Alzheimer's disease therefore provides an ideal tissue investigate incipient neurodegenerative changes. Here we have analyzed age‐ disease‐related occurrence composition GFAP human by using one‐ two‐dimensional electrophoresis, Western blots...

γ-Tubulin is assumed to participate in microtubule nucleation acentrosomal plant cells, but the underlying molecular mechanisms are still unknown. Here, we show that γ-tubulin present protein complexes of various sizes and different subcellular locations Arabidopsis fava bean. Immunoprecipitation experiments revealed an association with αβ-tubulin dimers. cosedimented microtubules polymerized vitro localized along their whole length. Large resistant salt treatment were found be associated a...

Profilin controls actin nucleation and assembly processes in eukaryotic cells. Actin elongation promoting factors (NEPFs) such as Ena/VASP, formins, WASP-family proteins recruit profilin:actin for filament formation. Some of these are found to be microtubule associated, making polymerization from microtubule-associated platforms possible. Microtubules implicated focal adhesion turnover, cell polarity establishment, migration, illustrating the coupling between systems. Here we demonstrate...

Abstract Background G protein-coupled receptor kinase-interacting proteins (GITs) function as GTPase-activating (GAPs) for small GTPases of the ADP-ribosylation factor (Arf) family. While GIT (GIT1 and GIT2) regulate both cell migration microtubule organization, their corresponding regulatory mechanisms in glioblastoma cells remain largely unknown. To further investigate role modulation, we examined GITs nucleation involvement protein kinase C (PKC) this process. Methods Glioblastoma lines...

ABSTRACT A protein of Mr 210 000 was identified in 3T3 cells by immunoblotting and immunoprecipitation with a monoclonal antibody MA-01. The thermo-labile located on microtubules prepared taxol-driven polymerization vitro. On fixed the MA-01 antigen interphase mitotic microtubular structures, vinblastine paracrystals, taxol bundles colcemid-resistant microtubules. Microin-jection experiments purified fol-lowed double immunofluorescence have shown that injection led to disruption vimentin...

A panel of 11 monoclonal antibodies specific to alpha- or beta-tubulin subunits was used study the location tubulin molecules in cytoplasmic microtubules. Specificity confirmed by immunoblotting and immunofluorescence experiments on fixed cells. The limited proteolysis with trypsin chymotrypsin followed demonstrated that discriminated between structural domains both subunits. Epitope mapping isolated alpha-tubulin revealed a set against N-terminal domain alpha-subunit (TU-01, TU-02, TU-03,...

Abstract Aggregation of the high-affinity IgE receptors (FcεRIs) on surface granulated mast cells initiates a chain signaling events culminating in release allergy mediators. Although microtubules are involved cell degranulation, molecular mechanism that controls microtubule rearrangement after FcεRI triggering is poorly understood. In this study, we show activation bone marrow-derived (BMMCs) induced by aggregation or treatment with pervanadate leads to rapid polymerization microtubules....

Abstract γ‐Tubulin is assumed to be a typical cytosolic protein necessary for nucleation of microtubules from microtubule organizing centers. Using immunolocalization and cell fractionation techniques in combination with siRNAi expression FLAG‐tagged constructs, we have obtained evidence that γ‐tubulin also present nucleoli mammalian interphase cells diverse cellular origins. Immunoelectron microscopy has revealed localization outside fibrillar centers where transcription ribosomal DNA takes...

Tubulin self-assembly into microtubules is a fascinating natural phenomenon. Its importance not just crucial for functional and structural biological processes, but it also serves as an inspiration synthetic nanomaterial innovations. The modulation of the tubulin process without introducing additional chemical inhibitors/promoters or stabilizers has remained elusive process. This work reports versatile vigorous strategy controlling by nanosecond electropulses (nsEPs). polymerization assessed...

Summary Many events during cell division are triggered by an evolutionary conserved regulator, the cyclin‐dependent kinase (Cdk). Here we used two novel drugs, purine analogues bohemine and roscovitine, to study role of Cdks in cycle progression microtubule organisation Vicia faba root tip cells. Both drugs inhibited activity immunopurified alfalfa Cdc2‐kinase. The transcript levels A‐ B‐type cyclin, as well cdc2 genes, declined treated tips, while mRNA level a D‐type cyclin gene was not...

Monoclonal antibodies were prepared against conserved synthetic peptide from the C-terminus of γ-tubulin and their specificity was confirmed by immunoblotting, competitive enzyme-linked immunosorbent assay (ELISA) immunofluorescence. The decorated interphase centrosomes as well half-spindles midbodies in mitotic cells various origin. used to study distribution nocodazole taxol-treated cells. In recovering treatment, found centers all microtubule asters. Examination relative location asters...

The molecular mechanisms controlling microtubule formation in cells with non-centrosomal microtubular arrays are not yet fully understood. key component of nucleation is γ-tubulin. Although previous results suggested that tyrosine kinases might serve as regulators γ-tubulin function, their exact roles remain enigmatic. In the present study, we show a pool associates detergent-resistant membranes differentiating P19 embryonal carcinoma cells, which exhibit elevated expression Src family...

Abstract In previous studies, we have shown overexpression and ectopic subcellular distribution of γ‐tubulin βIII‐tubulin in human glioblastomas glioblastoma cell lines (Katsetos et al., 2006, J Neuropathol Exp Neurol 65:455–467; Katsetos 2007, Neurochem Res 32:1387–1398). Here determined the expression surgically excised medulloblastomas (n = 20) medulloblastoma D283 Med DAOY. clinical tissue samples, immunohistochemical labeling was pervasive inversely related to neuritogenesis....

Abstract Activation of mast cells by aggregation the high-affinity IgE receptors (FcεRI) initiates signaling events leading to release inflammatory and allergic mediators stored in cytoplasmic granules. A key role this process play changes concentrations intracellular Ca2+ controlled store-operated entry (SOCE). Although microtubules are also involved degranulation, molecular mechanisms that control microtubule rearrangement during activation largely unknown. In study, we report bone...